Lipopolysaccharide modifies amiloride-sensitive Na+ transport processes across human airway cells

role of mitogen-activated protein kinases ERK 1/2 and 5

D. L. Baines, A. P. Albert, M. J. Hazell, Lorraine Gambling, A. M. Woollhead, M. E. C. Dockrell

    Research output: Contribution to journalArticle

    16 Citations (Scopus)

    Abstract

    Bacterial lipopolysaccharides (LPS) are potent inducers of proinflammatory signaling pathways via the activation of nuclear factor-kappa B (NF-kappa B) and mitogen-activated protein kinase (MAPK), causing changes in the processes that control lung fluid homeostasis and contributing to the pathogenesis of lung disease. In human H441 airway epithelial cells, incubation of cells with 15 mu g ml(-1) LPS caused a significant reduction in amiloride-sensitive I-sc from 15 +/- 2 to 8 +/- 2 mu A cm(-2) (p=0.01, n=13) and a shift in IC50 amiloride of currents from 6.8x10(-7) to 6.4x10(-6) M. This effect was associated with a decrease in the activity of 5 pS, highly Na+ selective, amiloride-sensitive < 1 mu M channels (HSC) and an increase in the activity of similar to 18 pS, nonselective, amiloride-sensitive > 10 mu M cation channels (NSC) in the apical membrane. LPS decreased alpha ENaC mRNA and protein abundance, inferring that LPS inhibited alpha ENaC gene expression. This correlated with the decrease in HSC activity, indicating that these channels, but not NSCs, were comprised of at least alpha ENaC protein. LPS increased NF-kappa B DNA binding activity and phosphorylation of extracellular signal-related kinase (ERK)1/2, but decreased phosphorylation of ERK5 in H441 cells. Pretreatment of monolayers with PD98059 (20 mu M) inhibited ERK1/2 phosphorylation, promoted phosphorylation of ERK5, increased alpha ENaC protein abundance, and reversed the effect of LPS on I-sc and the shift in amiloride sensitivity. Inhibitors of NF-kappa B activation were without effect. Taken together, our data indicate that LPS acts via ERK signaling pathways to decrease alpha ENaC transcription, reducing HSC/ENaC channel abundance, activity, and transepithelial Na+ transport in H441 airway epithelial cells.

    Original languageEnglish
    Pages (from-to)451-463
    Number of pages13
    JournalPflugers Archiv : European Journal of Physiology
    Volume459
    Issue number3
    Early online date13 Oct 2009
    DOIs
    Publication statusPublished - Feb 2010

    Keywords

    • Na+ channel
    • lung
    • airway
    • amiloride
    • LPS
    • ERK
    • epithelial sodium-channel
    • NF-Kappa-B
    • alpha-subunit
    • ENAC expression
    • gene-expression
    • down-regulation
    • glucocorticoid hormone
    • signaling pathway
    • ion-transport
    • camp
    • Na+ channel
    • Lung
    • Airway
    • Amiloride

    Cite this

    Lipopolysaccharide modifies amiloride-sensitive Na+ transport processes across human airway cells : role of mitogen-activated protein kinases ERK 1/2 and 5. / Baines, D. L.; Albert, A. P.; Hazell, M. J.; Gambling, Lorraine; Woollhead, A. M.; Dockrell, M. E. C.

    In: Pflugers Archiv : European Journal of Physiology, Vol. 459, No. 3, 02.2010, p. 451-463.

    Research output: Contribution to journalArticle

    Baines, D. L. ; Albert, A. P. ; Hazell, M. J. ; Gambling, Lorraine ; Woollhead, A. M. ; Dockrell, M. E. C. / Lipopolysaccharide modifies amiloride-sensitive Na+ transport processes across human airway cells : role of mitogen-activated protein kinases ERK 1/2 and 5. In: Pflugers Archiv : European Journal of Physiology. 2010 ; Vol. 459, No. 3. pp. 451-463.
    @article{b0c0c5e678dd4b3cae083f9082287465,
    title = "Lipopolysaccharide modifies amiloride-sensitive Na+ transport processes across human airway cells: role of mitogen-activated protein kinases ERK 1/2 and 5",
    abstract = "Bacterial lipopolysaccharides (LPS) are potent inducers of proinflammatory signaling pathways via the activation of nuclear factor-kappa B (NF-kappa B) and mitogen-activated protein kinase (MAPK), causing changes in the processes that control lung fluid homeostasis and contributing to the pathogenesis of lung disease. In human H441 airway epithelial cells, incubation of cells with 15 mu g ml(-1) LPS caused a significant reduction in amiloride-sensitive I-sc from 15 +/- 2 to 8 +/- 2 mu A cm(-2) (p=0.01, n=13) and a shift in IC50 amiloride of currents from 6.8x10(-7) to 6.4x10(-6) M. This effect was associated with a decrease in the activity of 5 pS, highly Na+ selective, amiloride-sensitive < 1 mu M channels (HSC) and an increase in the activity of similar to 18 pS, nonselective, amiloride-sensitive > 10 mu M cation channels (NSC) in the apical membrane. LPS decreased alpha ENaC mRNA and protein abundance, inferring that LPS inhibited alpha ENaC gene expression. This correlated with the decrease in HSC activity, indicating that these channels, but not NSCs, were comprised of at least alpha ENaC protein. LPS increased NF-kappa B DNA binding activity and phosphorylation of extracellular signal-related kinase (ERK)1/2, but decreased phosphorylation of ERK5 in H441 cells. Pretreatment of monolayers with PD98059 (20 mu M) inhibited ERK1/2 phosphorylation, promoted phosphorylation of ERK5, increased alpha ENaC protein abundance, and reversed the effect of LPS on I-sc and the shift in amiloride sensitivity. Inhibitors of NF-kappa B activation were without effect. Taken together, our data indicate that LPS acts via ERK signaling pathways to decrease alpha ENaC transcription, reducing HSC/ENaC channel abundance, activity, and transepithelial Na+ transport in H441 airway epithelial cells.",
    keywords = "Na+ channel, lung, airway, amiloride, LPS, ERK, epithelial sodium-channel, NF-Kappa-B, alpha-subunit, ENAC expression, gene-expression, down-regulation, glucocorticoid hormone, signaling pathway, ion-transport, camp, Na+ channel , Lung, Airway, Amiloride",
    author = "Baines, {D. L.} and Albert, {A. P.} and Hazell, {M. J.} and Lorraine Gambling and Woollhead, {A. M.} and Dockrell, {M. E. C.}",
    year = "2010",
    month = "2",
    doi = "10.1007/s00424-009-0717-4",
    language = "English",
    volume = "459",
    pages = "451--463",
    journal = "Pflugers Archiv : European Journal of Physiology",
    issn = "0031-6768",
    publisher = "Springer Verlag",
    number = "3",

    }

    TY - JOUR

    T1 - Lipopolysaccharide modifies amiloride-sensitive Na+ transport processes across human airway cells

    T2 - role of mitogen-activated protein kinases ERK 1/2 and 5

    AU - Baines, D. L.

    AU - Albert, A. P.

    AU - Hazell, M. J.

    AU - Gambling, Lorraine

    AU - Woollhead, A. M.

    AU - Dockrell, M. E. C.

    PY - 2010/2

    Y1 - 2010/2

    N2 - Bacterial lipopolysaccharides (LPS) are potent inducers of proinflammatory signaling pathways via the activation of nuclear factor-kappa B (NF-kappa B) and mitogen-activated protein kinase (MAPK), causing changes in the processes that control lung fluid homeostasis and contributing to the pathogenesis of lung disease. In human H441 airway epithelial cells, incubation of cells with 15 mu g ml(-1) LPS caused a significant reduction in amiloride-sensitive I-sc from 15 +/- 2 to 8 +/- 2 mu A cm(-2) (p=0.01, n=13) and a shift in IC50 amiloride of currents from 6.8x10(-7) to 6.4x10(-6) M. This effect was associated with a decrease in the activity of 5 pS, highly Na+ selective, amiloride-sensitive < 1 mu M channels (HSC) and an increase in the activity of similar to 18 pS, nonselective, amiloride-sensitive > 10 mu M cation channels (NSC) in the apical membrane. LPS decreased alpha ENaC mRNA and protein abundance, inferring that LPS inhibited alpha ENaC gene expression. This correlated with the decrease in HSC activity, indicating that these channels, but not NSCs, were comprised of at least alpha ENaC protein. LPS increased NF-kappa B DNA binding activity and phosphorylation of extracellular signal-related kinase (ERK)1/2, but decreased phosphorylation of ERK5 in H441 cells. Pretreatment of monolayers with PD98059 (20 mu M) inhibited ERK1/2 phosphorylation, promoted phosphorylation of ERK5, increased alpha ENaC protein abundance, and reversed the effect of LPS on I-sc and the shift in amiloride sensitivity. Inhibitors of NF-kappa B activation were without effect. Taken together, our data indicate that LPS acts via ERK signaling pathways to decrease alpha ENaC transcription, reducing HSC/ENaC channel abundance, activity, and transepithelial Na+ transport in H441 airway epithelial cells.

    AB - Bacterial lipopolysaccharides (LPS) are potent inducers of proinflammatory signaling pathways via the activation of nuclear factor-kappa B (NF-kappa B) and mitogen-activated protein kinase (MAPK), causing changes in the processes that control lung fluid homeostasis and contributing to the pathogenesis of lung disease. In human H441 airway epithelial cells, incubation of cells with 15 mu g ml(-1) LPS caused a significant reduction in amiloride-sensitive I-sc from 15 +/- 2 to 8 +/- 2 mu A cm(-2) (p=0.01, n=13) and a shift in IC50 amiloride of currents from 6.8x10(-7) to 6.4x10(-6) M. This effect was associated with a decrease in the activity of 5 pS, highly Na+ selective, amiloride-sensitive < 1 mu M channels (HSC) and an increase in the activity of similar to 18 pS, nonselective, amiloride-sensitive > 10 mu M cation channels (NSC) in the apical membrane. LPS decreased alpha ENaC mRNA and protein abundance, inferring that LPS inhibited alpha ENaC gene expression. This correlated with the decrease in HSC activity, indicating that these channels, but not NSCs, were comprised of at least alpha ENaC protein. LPS increased NF-kappa B DNA binding activity and phosphorylation of extracellular signal-related kinase (ERK)1/2, but decreased phosphorylation of ERK5 in H441 cells. Pretreatment of monolayers with PD98059 (20 mu M) inhibited ERK1/2 phosphorylation, promoted phosphorylation of ERK5, increased alpha ENaC protein abundance, and reversed the effect of LPS on I-sc and the shift in amiloride sensitivity. Inhibitors of NF-kappa B activation were without effect. Taken together, our data indicate that LPS acts via ERK signaling pathways to decrease alpha ENaC transcription, reducing HSC/ENaC channel abundance, activity, and transepithelial Na+ transport in H441 airway epithelial cells.

    KW - Na+ channel

    KW - lung

    KW - airway

    KW - amiloride

    KW - LPS

    KW - ERK

    KW - epithelial sodium-channel

    KW - NF-Kappa-B

    KW - alpha-subunit

    KW - ENAC expression

    KW - gene-expression

    KW - down-regulation

    KW - glucocorticoid hormone

    KW - signaling pathway

    KW - ion-transport

    KW - camp

    KW - Na+ channel

    KW - Lung

    KW - Airway

    KW - Amiloride

    U2 - 10.1007/s00424-009-0717-4

    DO - 10.1007/s00424-009-0717-4

    M3 - Article

    VL - 459

    SP - 451

    EP - 463

    JO - Pflugers Archiv : European Journal of Physiology

    JF - Pflugers Archiv : European Journal of Physiology

    SN - 0031-6768

    IS - 3

    ER -